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Chapter 37 : Molecular Comparison of Isolates from a Recurring Outbreak of Legionnaires’ Disease Spanning 22 Years

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Molecular Comparison of Isolates from a Recurring Outbreak of Legionnaires’ Disease Spanning 22 Years, Page 1 of 2

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Abstract:

During outbreak investigations molecular analytical methods are necessary to discriminate among clinical isolates and to establish a link with environmental isolates. In addition to monoclonal antibodies used as an initial subtyping method, other methods employed have been pulse-field gel electrophoresis, amplified fragment length polymorphosim (AFLP), arbitrarily primed PCR, multilocus variable-number tandem repeat, and sequence-based typing (SBT). In the present study, the authors used SBT to study the relationships among clinical and environmental isolates of isolated from the same hotel during a 22-year period. Water samples were collected from the hotel and cultured using standard procedures for the recovery of isolates from environmental sources. To determine the discriminatory power of the SBT procedure, eight isolates that share the same monoclonal antibody (MAb) subtype were included. These isolates represent four enzyme types based on previous studies using multilocus enzyme electrophoresis (MLEE). All isolates were tested with a panel of MAbs to determine their subtype. Isolates were compared by AFLP. Patient isolates and representative environmental isolates from both outbreaks were analyzed by a sequence-based typing scheme.

Citation: F. Benson R, E. Lucas C, W. Brown E, D. Cowgill K, S. Fields B. 2006. Molecular Comparison of Isolates from a Recurring Outbreak of Legionnaires’ Disease Spanning 22 Years, p 139-142. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch37

Key Concept Ranking

Multilocus Sequence Typing
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Legionella pneumophila
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Figures

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FIGURE 1

Genetic relatedness of St. Croix patient isolates from 1981 and 2003 and reference serogroup 1 MAb 1,2,3 pattern.

Citation: F. Benson R, E. Lucas C, W. Brown E, D. Cowgill K, S. Fields B. 2006. Molecular Comparison of Isolates from a Recurring Outbreak of Legionnaires’ Disease Spanning 22 Years, p 139-142. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch37
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References

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1. Aurell, H.,, J. Etienne,, F. Forey,, M. Reyrolle,, P. Girardo,, P. Farge,, B. Decludt,, C. Campese,, F. Vandenesch, and, S. Jarraud. 2003. Legionella pneumophila serogroup 1 strain Paris: endemic distribution throughout France. J. Clin. Microbiol. 41:33203322.
2. Aurell, H.,, P. Farge,, H. Meugnier,, M. Gouy,, F. Foey,, G. Lina,, F. Vandenesch,, J. Etienne, and, S. Jarraud. 2005. Clinical and environmental isolates of Legionella pneumophila serogroup 1 cannot be distinguished by sequence analysis of two surface protein genes and three housekeeping genes. Appl. Environ. Microbiol. 71:282289.
3. Benson, R. F.,, B. S. Fields,, A. Benin,, H. Craddock, and, R. E. Besser. 2002.Application of amplified fragment length polymorphism analysis to subtyping of Legionella pneumophila serogroup 6, p. 243247. In R. Marre,, Y. Abu Kwaik,, C. Bartlett,, N. P. Cianciotto,, B. S. Fields,, M. Frosch,, J. Hacker,, P. C. Luck (ed.). Legionella. ASM Press, Washington, D.C.
4. Darelid, J.,, S. Bernander,, K. Jacobson, and, S. Lofgren. 2004. The presence of a specific genotype of Legionella pneumophila serogroup 1 in a hospital and municipal water distribution system over a 12-year period. Scand. J. Infect. Dis. 36:417423.
5. Gaia, V.,, N. K. Fry,, B. Afshar,, P. C. Luck, et al. 2005. Consensus sequence-based scheme for epidemiological typing of clinical and environmental isolates of Legionella pneumophila. J. Clin. Microbiol. 43:20472052.
6. Kumar, S.,, K. Tamura, and, M. Nei. 2004. MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment. Briefings Bioinformatics 5:150163.
7. Pruckler, J. M.,, L. A. Mermel,, R. F. Benson,, C. Giorgio,, P. K. Cassiday,, R. F. Breiman,, C. G. Whitney, and, B. S. Fields. 1995. Comparison of Legionella pneumophila isolates by arbitrarily primed PCR and pulsed-field gel electrophoresis: analysis from seven epidemic investigations. J. Clin. Microbiol. 33:28722875.
8. Rangel-Frausto, M. S.,, P. Rhomberg,, R. J. Hollis,, M. A. Pfaller,, R. P. Wenzel,, C. M. Helms, and, L. A. Herwaldt. 1999. Persistence of Legionella pneumophila in a hospital’s water system: a 13-year survey. Infect. Control. Hosp. Epidemiol. 20:793797.
9. Selander, R. K.,, R. M. McKinney,, T. S. Whittam, W. F. Bibb,, D. J. Brenner,, F. S. Nolte, and, P. E. Pattison. 1985. Genetic structure of populations of Legionella pneumophila J. Bacteriol. 163:10211037.

Tables

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TABLE 1

SBT for isolates obtained from St. Croix, U.S.Virgin Islands during 1981 and 2003 epidemic investigations and reference strains

Citation: F. Benson R, E. Lucas C, W. Brown E, D. Cowgill K, S. Fields B. 2006. Molecular Comparison of Isolates from a Recurring Outbreak of Legionnaires’ Disease Spanning 22 Years, p 139-142. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch37

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